脊尾白虾14-3-3 基因cDNA 全长的克隆和表达分析

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脊尾白虾14-3-3 基因cDNA 全长的克隆和表达分析
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                        王有昆1王有昆1
1. 上海海洋大学 水产与生命学院, 上海 201306; 2. 农业部海洋渔业可持续发展重点实验室, 中国水产科学研究院 黄海水产研究所, 山东 青岛 266071; 3. 青岛海洋科学与技术国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266235
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1. 上海海洋大学 水产与生命学院, 上海 201306; 2. 农业部海洋渔业可持续发展重点实验室, 中国水产科学研究院 黄海水产研究所, 山东 青岛 266071; 3. 青岛海洋科学与技术国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266235
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刘萍2刘萍2
1. 上海海洋大学 水产与生命学院, 上海 201306; 2. 农业部海洋渔业可持续发展重点实验室, 中国水产科学研究院 黄海水产研究所, 山东 青岛 266071; 3. 青岛海洋科学与技术国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266235
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1. 上海海洋大学 水产与生命学院, 上海 201306; 2. 农业部海洋渔业可持续发展重点实验室, 中国水产科学研究院 黄海水产研究所, 山东 青岛 266071; 3. 青岛海洋科学与技术国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266235
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李吉涛2李吉涛2
1. 上海海洋大学 水产与生命学院, 上海 201306; 2. 农业部海洋渔业可持续发展重点实验室, 中国水产科学研究院 黄海水产研究所, 山东 青岛 266071; 3. 青岛海洋科学与技术国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266235
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李健2李健2
1. 上海海洋大学 水产与生命学院, 上海 201306; 2. 农业部海洋渔业可持续发展重点实验室, 中国水产科学研究院 黄海水产研究所, 山东 青岛 266071; 3. 青岛海洋科学与技术国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266235
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作者单位:1. 上海海洋大学 水产与生命学院, 上海 201306; 2. 农业部海洋渔业可持续发展重点实验室, 中国水产科学研究院 黄海水产研究所, 山东 青岛 266071; 3. 青岛海洋科学与技术国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266235
作者简介:作者简介： 王有昆(1989–), 硕士, 主要从事甲壳类分子辅助育种研究. E-mail： wangyoukun1989@sina.com 通信作者：刘萍, 硕士生导师, 研究员. E-mail： liuping@ysfri.ac.cn
通讯作者:
中图分类号:S917
基金项目:国家863 计划项目(2012AA10A409)； 国家虾产业技术体系项目(CARS-47)；青岛海洋科学与技术国家实验室鳌山科技创新项目(2015ASKJ02).

Cloning and expression analysis of 14-3-3 gene from Exopalaemon carinicauda

Author:

WANG Youkun1
WANG Youkun1
1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China; 2. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sci
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2
1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China; 2. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sci
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LIU Ping2
LIU Ping2
1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China; 2. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sci
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3
1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China; 2. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sci
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LI Jitao2
LI Jitao2
1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China; 2. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sci
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LI Jian2
LI Jian2
1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China; 2. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sci
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Affiliation:

1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China; 2. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sci

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摘要:

利用本实验室构建的脊尾白虾(Exopalaemon carinicauda)血细胞cDNA 文库中脊尾白虾14-3-3 巨球蛋白基因EST 序列, 采用cDNA 末端快速扩增(RACE)技术, 克隆获得脊尾白虾14-3-3 基因cDNA 全长, 命名为Ec14-3-3。该基因全长2905 bp, 包含744 bp 的开放阅读框, 编码247 个氨基酸组成的蛋白质, 分子量为27.95 kD, 理论等电点为4.65。同源性分析表明, 脊尾白虾Ec14-3-3 氨基酸序列与斑节对虾(Penaeus monodon) 14-3-3 的同源性最高, 达到98%。荧光定量PCR 分析结果表明, Ec14-3-3 基因在血细胞、卵巢、肝胰腺等组织中均有表达, 其中血细胞中Ec14-3-3相对表达量最高。感染鳗弧菌和WSSV 后6 h, 脊尾白虾血细胞和肝胰腺中Ec14-3-3 基因的表达量较对照组均显著增加(P<0.05), 且具有明显的时间差异性。本研究结果表明, Ec14-3-3 基因在脊尾白虾免疫反应中发挥重要作用。

关键词:脊尾白虾; 14-3-3; 基因克隆; 基因表达

Abstract:

Exopalaemon carinicauda is the most commercially important shrimp in China based on reproduction, along growing season, and ecological flexibility. However, the frequency of disease outbreaks has increased asproduction has increased, resulting in huge economic losses. E. carinicauda relies on the innate immune responseto defend against invading pathogens. The 14-3-3 family of proteins is highly conserved and present in all eukaryoticcells. The 14-3-3 proteins are involved in many cellular pathways, including those for metabolism, thecell cycle, differentiation, signaling, apoptosis, and neoplastic transformation. Based on the 14-3-3 expressed sequencetag from a hemocyte cDNA library of E. carinicauda, the full-length cDNA of 14-3-3 from E. carinicauda(named Ec14-3-3) was cloned using the rapid amplification of cDNA ends technique. The Ec14-3-3 cDNA was2 905 bp in length and contained an open reading frame of 744 bp, encoding a mature protein of 247 amino acids,with a molecular mass of 27.95 kD and an estimated pI of 4.65. Homology analysis revealed that the Ec14-3-3amino acid sequence was highly conserved with homologs from other crustaceans, and amino acid sequence similaritywas 98% with the 14-3-3 from Penaeus monodon. Ec14-3-3 expression levels were measured in differenttissues by quantitative real-time polymerase chain reaction analysis. Ec14-3-3 expression was detected in hemocytes,gill, hepatopancreas, muscle, ovary, intestine, stomach, and the eyestalk of E. carinicauda, with the highestexpression level in hemocytes. Ec14-3-3 expression was upregulated in hemocytes and the hepatopancreas afterchallenges with Vibrio anguillarum and white spot syndrome virus. These results suggest that Ec14-3-3 may playan important role in the prawn immune response.

Key words:Exopalaemon carinicauda; 14-3-3 gene; gene cloning; gene expression

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引用本文

王有昆,刘萍,李吉涛,李健.脊尾白虾14-3-3 基因cDNA 全长的克隆和表达分析[J].中国水产科学,2016,23(1):44-52
WANG Youkun, LIU Ping, LI Jitao, LI Jian. Cloning and expression analysis of 14-3-3 gene from Exopalaemon carinicauda[J]. Journal of Fishery Sciences of China,2016,23(1):44-52

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